研究血管抑素 (angiostatin)的临床应用价值 ,将其开发为多肽类药物。方法 :以RT PCR从胎儿肝脏钓取人血管抑素全编码区cDNA ,使用Pichia分泌型酵母表达系统表达重组人血管抑素 ,重组蛋白经肝素亲和层析纯化后 ,以鸡胚尿囊膜法血管生成实验及小鼠创伤愈合实验检测活性。结果 :重组人血管抑素在酵母系统中得到较高量表达 (5mg/L) ,经肝素亲和层析纯化 ,SDS PAGE显示分子量为 43kD。活性实验表明 ,重组蛋白能够抑制新生血管形成、抑制伤口愈合。结论 :重组人血管抑素在酵母系统中实现高效表达 ,并具有很好的生物学活性。

Objective: To express recombinant human angiostatin for further application in clinic. Methods: The complete encoding cDNA of human angiostatin was isolated from human embryo liver with RT PCR and expressed in secretory Pichia expression system. Recombinant human angiostatin was purified with heparin sepharose chromatography and its activity was determined in chick embryo chorioallantoic membrane (CAM) and wound healing assays. Results: Expressed in large quantity (yield=5 mg/L) and purified with heparin sepharose, recombinant angiostatin was showed to have a molecular weight of 43 kD in SDS PAGE and potently inhibit angiogenesis and wound healing. Conclusion: Recombinant human angiostatin was expressed efficiently in a biologically active form.